In vitro transcytosis assay

Abstract

The present disclosure relates to methods and compositions useful for measuring the transcytosis of a molecule. In particular, the present disclosure relates to in vitro receptor-dependent transcytosis assays for evaluating the clearance rates of therapeutic antibody molecules and Fc-fusion proteins in humans and animals.

What is claimed is: 
     
       1. A method of determining the in vivo clearance of a monoclonal antibody, comprising:
 a) introducing the monoclonal antibody into a first of two chambers where the first chamber is separated from the second chamber by a monolayer of Madin-Darby canine kidney (MDCK) cells expressing a human neonatal Fc receptor (FcRn) heavy chain and a human beta-2-microglobulin, and wherein each of the first and second chambers has a physiological pH value of 7.4; 
 b) measuring the number of monoclonal antibodies transcytosed from the first chamber to the second chamber; and 
 c) determining the in vivo clearance of the monoclonal antibody based on the number of monoclonal antibodies measured. 
 
     
     
       2. The method of  claim 1 , wherein measuring the number of the monoclonal antibodies transcytosed comprises use of an enzyme-linked immunosorbent assay (ELISA), liquid-scintillation counting (LSC), quantitative PCR, or a fluorescence reader system.